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Additional file 1: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Figure S1. LSTM microscopy implementation. (a) Image of the physical LSTM setup. (b) 3D model of LSTM and the sample mounting system. The 3D-printed sample chamber is designed to accommodate large biological samples of virtually any dimensions, while still allowing the objectives to be immersed in the immersion oil. Two transparent glass windows, located on the lateral sides, provide visual view of the sample for ease of positioning. An additional window is realized at the bottom part of the chamber to allow the illumination light to pass through. An additional adapter was designed to allow mounting a prism mirror at about approximately 10° from the normal surface to facilitate the optical alignment of the system. (PDF 1623 kb

Additional file 9: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Video 4. High-resolution LSTM imaging of a large tissue of Thy1-eYFP mouse brain. The bounding box is 9.6 mm × 13.5 mm × 5.34 mm. The raw data was down-sampled 4 × 4 fold to make the volume rendering feasible. The high-resolution video is available in the figshare repository at https://doi.org/10.6084/m9.figshare.c.4072160 . (MOV 167936 kb

Additional file 10: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Video 5. Visualization of an image stack of vasculature-stained rat brain tissue. This video visualizes an image stack acquired from a large rat brain slice (stained for vasculature with tomato lectin) using LSTM in 2-AS mode. The bounding box is 1 mm × 1 mm × 5 mm. The high-resolution video is available in the figshare repository at https://doi.org/10.6084/m9.figshare.c.4072160 . (MOV 143360 kb

Additional file 13: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Video 8. Neuronal activity traces of representative neurons. A visualization of the neuronal traces shown in Fig. 7b. The high-resolution video is available in the figshare repository at https://doi.org/10.6084/m9.figshare.c.4072160 . (MOV 11161 kb

Additional file 5: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Video 2. Comparison of image volumes acquired with LSTM in 1-axis scan (1-AS) and 2-axis scan (2-AS) modes. The 3D rendering visualizes the image stacks acquired from the same sample (human brain section stained with DAPI) with LSTM in 1-AS and simultaneous 2-AS modes. The high-resolution video is available in the figshare repository at https://doi.org/10.6084/m9.figshare.c.4072160 . (MOV 104448 kb

Additional file 11: of Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Video 6. High-resolution LSTM imaging of a large expanded section of Thy1-eYFP mouse brain. A thin (250 μm) coronal section was expanded ~ 4-fold using proExM procedure and imaged using LSTM with 10×/0.6NA detection objective. The resulting dataset (~ 6 TB) consisted of 723,300 full frame images (2048 × 2048). The data was down-sampled 8 × 8 fold to allow high-quality volumetric rendering. The high-resolution video is available in the figshare repository at https://doi.org/10.6084/m9.figshare.c.4072160 . (MOV 439296 kb

Video 8: Neuronal activity traces of representative neurons.

A visualization of the neuronal traces shown in Figure 7b of the related publication; presenting rapid volumetric calcium imaging of highly motile Hydra.<div><br><div>An advantage of the LSTM imaging technique (see below for details) is the capacity for rapid volumetric live imaging of samples undergoing substantial non-isomorphic rearrangements in their body shape and cellular density, resulting in continuously changing local optical properties.<br><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div></div

Video 2: Comparison of image volumes acquired with LSTM in 1-AS and 2-AS modes.

<div><p>The 3D rendering visualizes the image stacks acquired from the same sample (human brain section stained with DAPI) with LSTM in 1-Axis Scan (1-AS) and simultaneous 2-Axes Scan (2-AS) mode.</p><p><br></p> <p>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</p></div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p

Video 7: Rapid volumetric calcium imaging of highly motile Hydra.

GCaMP6s expressing Hydra was imaged using LSTM with 10x/0.6NA objective. Maximum intensity projections are shown for the two halves of the volume. First occurrences of longitudinal and radial contractions are annotated. The scale Bar is 100 microns.<div><br></div><div>An advantage of the LSTM imaging technique (see below for details) is the capacity for rapid volumetric live imaging of samples undergoing substantial non-isomorphic rearrangements in their body shape and cellular density, resulting in continuously changing local optical properties.<div><div><br></div><div>This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.</div><div><br></div><div>LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.</div><div><br></div><div>View the full collection of related videos at: </div><p><a href="https://doi.org/10.6084/m9.figshare.c.4072160">https://doi.org/10.6084/m9.figshare.c.4072160</a></p></div></div